Abstract

This unit describes two methods of in situ hybridization: one uses an 35S‐labeled oligonucleotide probe and the other uses a digoxigenin‐labeled oligonucleotide probe on frozen, cryostat‐sectioned
samples. These methods allow detection of the physical distribution and expression levels of target mRNA. Protocols are also
included for labeling the probes and preparing the sample material.

CAUTION: When working with radioactivity, take appropriate precautions to avoid contamination of the experimenter and surroundings.
Carry out the experiment and dispose of wastes in an appropriately designated area following the guidelines provided by your
local radiation safety offices (also see appendix 1A).

CAUTION: When working with radioactivity, take appropriate precautions to avoid contamination of the experimenter and surroundings.
Carry out the experiment and dispose of wastes in an appropriately designated area following the guidelines provided by your
local radiation safety offices (also see appendix 1A).

Support Protocol 2: 3′ Labeling of Oligonucleotide Probes with Digoxigenin